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Gladstone scientists develop technique to decipher the dormant AIDS virus concealed in cells
Date:9/11/2012

acking the individual cells that traditionally had been difficult to monitor."

Singe-cell, time-lapse microscopya state-of-the-art technique that scientists have lately used to track some viral infections and map antibiotic resistance to drugshas not worked for tracking the HIV-infection cycle in CD4 T cells, especially in the latent state. This is because these cells are notoriously evasive. They spontaneously move around, attaching and detaching from their neighbors, making it nearly impossible to monitor individual HIV-infected cells over time.

However, Dr. Weinberger's team devised a clever system that essentially guides and suspends HIV-infected T cells into tiny finger-like channelsreducing their ability to move or detach from their neighbors.

"First, we load the T cells into a small well, allowing them to settle into the bottomwhich is filled with nutrients that keep the cells well-fed and stress-free," explained the paper's lead author Brandon Razooky, a Gladstone and UCSF graduate student. "Next, we tilt the device and the cells slide into microscopic finger-like channels that are attached to the well. Finally, we return the device to its upright position, locking about 25 T cells inside each channel and essentially 'freezing' them in place."

The device has several advantages over current methods. First and foremost, individual cells stay in place so investigators can follow them over time with single-cell, time-lapse microscopy. Second, the fact that each T cell is suspended in nutrients in close physical contact with other cells results in near optimal conditions for keeping the infected cell alive for the virus' entire life cycle.

"This means that we now have the potential to analyze the entire course of an HIV infection in an individual cellespecially during the crucial latency stagefor which we know so little," said Dr. Weinberger. "In the future, we plan to expand the device's design to include a larger nu
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Contact: Anne Holden
anne.holden@gladstone.ucsf.edu
415-734-2534
Gladstone Institutes
Source:Eurekalert  

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